Eph and Lyn hyper-phosphorylation and CRMP interactions in AD"

AD中Eph和Lyn过度磷酸化与CRMP相互作用"

基本信息

批准号：
10746170
负责人：
MATTHIAS BUCK
金额：
$ 24.15万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-15 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10746170
关键词：
Actins Affinity Alzheimer&apos s Disease Amyloid beta-Protein Antibodies Antigens Binding Biochemical Bioinformatics Biophysics Brain C-terminal Cardiovascular system Cells Complex Computer Models Cytoskeleton Data Development Diagnostic Disease Docking Early Diagnosis Eph Family Receptors Exploratory/Developmental Grant Family Family member Future Future Generations Grain Homologous Gene In Vitro Knowledge Learning Link Mammalian Cell Mediator Memory Microtubules Molecular Mutagenesis Mutation Natural regeneration Nature Neurodegenerative Disorders Neuronal Injury Neurons Organism Parkinson Disease Pattern Peptides Phosphorylation Phosphotransferases Preparation Process Protein Kinase Proteins Research Research Personnel Risk Role SAM Domain Sampling Semaphorin-3A Site Structure System Therapeutic Therapeutic Intervention Time Validation aging brain antibody diagnostic biophysical techniques cell motility design experimental study follow-up glycogen synthase kinase 3 beta insight mimetics molecular dynamics molecular modeling neuron development plexin potential biomarker protein complex protein purification reconstitution response scaffold structural biology targeted treatment tau Proteins tool

项目摘要

Eph receptors make guidance decisions for cell migration in cardiovascular and neuronal development, disease, and regeneration. Eph receptors are also involved with higher brain functions, memory and learning. A protein that has been associated with Alzheimer’s, Parkinson’s and other neuronal diseases and injuries is the Collapsin Response Mediator Protein (CRMP) which interacts with several kinases and becomes hyper-phosphorylated alongside increased activation of the kinases. The hyper- phosphorylated CRMP then disrupts the formation of actin and microtubule cytoskeletal structures and it thought to impede Aβ and tau clearance. Cytosolic Lyn kinase, a close homologue of Fyn kinase, is known to interact directly with EphA4 and here, for the first time, we show that an EphA family member and EphB2 directly interacts with CRMP. Analogous to another guidance and cell migration system, the Fyn-Plexin-CRMP complex, the Lyn-EphA4-CRMP association is likely to form a complex with Cdk5 and GSK3β kinases, each also involved in Alzheimer’s. The features of the Eph-CRMP and Eph-Lyn interacting interfaces need to be characterized, as they will be potential biomarkers and targets for therapeutic intervention. The proposal has two main aims. Aim 1) seeks to establish the in vitro phosphorylation patters of various kinases on the intracellular domains of EphA4 and –B2 in the presence and absence of CRMP and to further validate the formation of the complex in cells. Aim 2) The effect that the corresponding phosphomimetic/phospho-defective mutations have on the level of activity on these Eph receptor intracellular regions and of the kinases will be studied with purified proteins in vitro. Eventually, using the knowledge from this project, Antibodies or complex-disrupting-peptides may drive the future development of early detection diagnostics and/or therapeutics.

Eph受体为心血管和神经元发育中的细胞迁移做出指导决策， Eph 受体还与高级脑功能、记忆力和再生有关。一种与阿尔茨海默病、帕金森病和其他神经元疾病有关的蛋白质。损伤是与多种激酶相互作用的崩溃蛋白反应介导蛋白 (CRMP) 并随着激酶激活的增加而变得过度磷酸化。磷酸化的 CRMP 会破坏肌动蛋白和微管细胞骨架结构的形成，细胞溶质 Lyn 激酶（Fyn 激酶的密切同源物）被认为会阻碍 Aβ 和 tau 清除。已知与 EphA4 直接相互作用，在这里，我们首次证明 EphA 家族成员 EphB2 直接与 CRMP 相互作用，类似于另一个引导和细胞迁移系统， Fyn-Plexin-CRMP 复合物，Lyn-EphA4-CRMP 关联可能与 Cdk5 形成复合物， GSK3β 激酶，每种都与阿尔茨海默病有关 Eph-CRMP 和 Eph-Lyn 的特征。相互作用的界面需要进行表征，因为它们将是潜在的生物标志物和目标该提案有两个主要治疗目标 1) 寻求建立体外治疗。各种激酶在 EphA4 和 –B2 细胞内结构域上的磷酸化模式 CRMP 的存在和不存在并进一步验证细胞中复合物的形成。相应的磷酸模拟/磷酸缺陷突变对活性水平的影响将使用纯化的蛋白质来研究这些 Eph 受体胞内区域和激酶的作用最终，利用该项目的知识，抗体或复合物破坏肽可能会产生。推动早期检测诊断和/或治疗的未来发展。